I need to use RNAse protection to evaluate message levels in transgenic
mice. I'm starting from scratch with the chapter from "Current Protocols
in Mol. Biol." and the Ambion catalog. Answers to any of the following
questions will be greatly appreciated.
Is Ambion's RPA-II kit worth the money?
Does anyone out there have a reliable protocol and/or friendly advice for a
beginner?
Is it worth buying template for size markers? $75 for 5 ug seems a bit
steep.
Does anyone have a plasmid containing a mouse housekeeping gene cDNA that
I could use as a probe template for positive control/quantitation?
Who makes the best kit for making probes? Can I get by with ordering new
polymerases and using the buffers, rNTPs, etc. that I've used in the past
for riboprobes?
Would it be worthwhile to begin by using RNA transcribed in vitro from my
transgene constructs?
Thank you!
Merideth Kelley
mlkelley at u.washington.edu